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Microenvironmental Analysis and Control for Local Cells under Confluent Conditions via a Capillary-Based Microfluidic
Nobutoshi Ota1, Nobuyuki Tanaka1, Asako Sato1
1Center for Biosystems Dynamics Research (BDR), RIKEN, 1-3 Yamadaoka, Suita, Osaka565-0871, Japan.
Analytical Chemistry
|November 16, 2022
Summary
This study introduces a new microfluidic device for analyzing cell microenvironments in culture dishes. It enables localized cell stimulation and molecule collection from confluent cell cultures, advancing cellular microenvironment research.
Area of Science:
- Cell Biology
- Microfluidics
- Tissue Engineering
Background:
- Cellular microenvironments are crucial for tissue function and are influenced by secreted molecules.
- Existing microfluidic devices are unsuitable for analyzing cells in confluent culture conditions.
- Studying confluent cells is essential for understanding typical cell and tissue cultivation setups.
Purpose of the Study:
- To develop a novel microfluidic device for studying confluent cells in culture dishes.
- To enable localized cell stimulation and simultaneous collection of secreted molecules.
- To analyze the cellular microenvironment under physiologically relevant conditions.
Main Methods:
- A glass capillary-based microfluidic device was designed to create confined local flow (≤100 μm scale).
- The device facilitated simultaneous local cell stimulation and secretion collection in adjacent chemical regions.
- Experiments utilized mouse skeletal myoblast (C2C12) and leukemic monocyte (RAW264) cell lines under confluent conditions.
Main Results:
- Localized cell removal (7.6 ± 2.0 cells) was achieved using trypsin stimulation.
- Tumor necrosis factor alpha (TNF-α) secretion from stimulated RAW264 cells was approximately four-fold higher than unstimulated cells within 90 minutes.
- The device successfully enabled microenvironmental analysis of stimulated and unstimulated cells.
Conclusions:
- The novel microfluidic device effectively supports the study of confluent cell cultures.
- It allows for precise local stimulation and collection of secreted molecules, crucial for microenvironment analysis.
- This technology advances research into cellular behavior and function in confluent conditions.
